IL-33 Mediates Lung Inflammation by the IL-6-Type Cytokine Oncostatin M

Figure 9

Comparison of AdOSM-induced IL-33 expression in C57Bl/6 mouse lung using the Nessy-1 monoclonal antibody and OSM-induced IL-33 upregulation in Human Type 2 Alveolar Epithelial Cells (A549). (a) C57Bl/6 wild-type (WT) or IL-33 knockout (IL-33KO) mice were endotracheally administered AdDl70 (control) or AdOSM for 7 days, and whole lung cell lysate was analyzed for the expression of IL-33 by Western blotting using the Nessy-1 monoclonal antibody. (b) 300,000 A549 cells/well in 6-well plates were stimulated with human OSM (20 ng/ml) for 24 hours, and Western blot analysis was performed for detection of IL-33 (Nessy-1), pSTAT3 (NEB Cell Signaling), tSTAT3 (NEB Signaling), and actin (Santa Cruz) from cell lysates. Black arrow indicates migration of IL-33-specific band. Mouse IgG light chain. (c) Densitometry of IL-33 and pSTAT3/tSTAT3 signal was completed using Image Studio Lite for samples in (b). AU: arbitrary units.

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